翻译组图谱测序技术的优化方法的评估

Ribosome profiling (Ribo-seq) is a high-throughput sequencing technique that captures the translational state of mRNAs at codon-level resolution. It has been widely applied to study translation efficiency, non-canonical translation events, and stress response mechanisms. However, the applicability of this technique across different species and sample types remains limited by library construction strategies and the choice of key nucleases. To enhance the generalizability and data quality of Ribo-seq, this study focused on optimizing the nuclease digestion step and systematically evaluated the performance of nuclease P1, recently shown to be effective in mammalian cells and yeast, in Ribo-seq experiments. The quality of Drosophila Ribo-seq libraries generated using nuclease P1 was assessed. Quality control and data filtering of the constructed libraries yielded over 900,000 high-quality reads, ensuring data reliability and analytical feasibility. This study demonstrates the suitability of nuclease P1 in Ribo-seq experiments, providing a robust data foundation for future studies on translational regulation and offering a reproducible technical framework for translatomic research across species.